强直性脊柱炎滑膜组织破骨细胞表型、TNF-α及MMP-3的表达及对软骨破坏作用的研究

THE PHENOTYPE OF OSTEOCLAST,EXPRESSION OF TNF-α AND MMP-3 IN SYNOVIUM OF ANKYLOSING SPONDYLITIS AND THEIR DESTRUCTIVE EFFECT ON CARTILAGE

目的检测破骨细胞表型、基质金属蛋白酶-3及肿瘤坏死因子-α在强直性脊柱炎患者滑膜组织的表达,滑膜单核巨噬细胞与软骨共培养,了解其在强直性脊柱炎软骨破坏中的作用,探讨强直性脊柱炎软骨破坏机制。方法采用酶组织化学技术检测滑膜组织内破骨细胞表型表达,原位杂交技术检测强直性脊柱炎患者滑膜组织内肿瘤坏死因子-α、基质金属蛋白酶-3的表达,LeicaQwin高清晰图像分析系统测定阳性细胞数,强直性脊柱炎滑膜单核巨噬细胞培养并与正常软骨共培养,HE染色及扫描电镜观察软骨的破坏程度。结果破骨表型细胞在滑膜组织内阳性表达,阳性细胞计数与对照组有显著差异(P<0.01);肿瘤坏死因子-α、基质金属蛋白酶-3在强直性脊柱炎患者滑膜细胞内有棕黄色颗粒沉着,阳性细胞计数与对照组有显著差异(P<0.01);滑膜细胞与正常软骨共培养,HE染色见软骨表面粗糙,有小凹陷,扫描电镜可见软骨表面不定形物质成斑片状,胶原纤维暴露,有断裂,抗肿瘤坏死因子-α可以减轻软骨的破坏程度。结论破骨表型细胞、肿瘤坏死因子-α及基质金属蛋白酶-3是软骨破坏的重要因素,破骨表型细胞通过表达基质金属蛋白酶-3分解软骨基质,进而发挥软骨的破坏作用,抗肿瘤坏死因子-α可以抑制破骨表型细胞对软骨的破坏。

Objective To detect the expression of the osteoclast phenotype, matrix metalloproteinase-3 and tumor necrosis factor-α in synovium tissue of ankylosing spondylitis, and to understand the effects and the mechanism of the mononuclear macrophage in the cartilaginous damage of ankylosing spondylitis by co-culturing synovium mononuclear macrophages and normal catrilage. Methods We used enzymohistochemisty and in-situ hybridization respectively to detect the expression of the osteoclast phenotype, MMP-3 and TNF-α in the synovium of ankylosing spondylitis, used Leica Qwin analytical system to determine the number of positive cells; cocultured synoviocytes and cartilage, and observed the degree of cartilaginous damage with HE staining and scanning electron microscopy. Results 1. Synoviocytes with the osteoclast phenotype existed generally in the synovium of AS, and their number was significantly greater than that in the normal synovium （P〈 0.01）. 2. MMP-3 and TNF-α were expressed strongly in the synoviocytes of AS, also significantly more than those in the normal synovium （P〈 0.01）. 3. In cocultured cartilage with mononuclear macrophages, HE stainning and scanning electron microscopy, rerealed exposure and collapse of the collagen at the surface of cartilage, and anti-TNF-α could relieve the degree of destruction of cartilage. Conclusion Osteoclast phenotype cells, TNF-α and MMP-3 are important factors of cartilage dustruction. Osteoclast phenotype cells may decompose cartiage matrix by expressing MMP-3 to destruct cartilage, but anti-TNF-α may inhibit cartilage decomposion.